LOCUS       40924_24298        8856 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHC93, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8856)
  AUTHORS   Chou H-H., Berthet J, Marx CJ.
  TITLE     Growth rate modulates the selective advantage of recurrent 
            transposition mutations arising from experimental populations grown 
            in a metal-limited environment
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8856)
  AUTHORS   Chou H-H.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-OCT-2008) Organismic and Evolutionary Biology, Harvard
            University, 16 Divinity Avenue, Biolab 3079, Cambridge, MA 02138, 
            USA
REFERENCE   3  (bases 1 to 8856)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8856)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (15-OCT-2008) Organismic and Evolutionary Biology, Harvard 
            University, 16 Divinity Avenue, Biolab 3079, Cambridge, MA 02138, 
            USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8856
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      3..712
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      1105..1693
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    1981..2002
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2017..2047
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2055..2071
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2079..2095
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(2133..2864)
                     /codon_start=1
                     /gene="icuB"
                     /product="IcuB"
                     /label=icuB
                     /protein_id="ACR44983.1"
                     /translation="MRRTVILTALLAFTAPAAAHDLWLDPAGNGVQILYGHPHEPELPS
                     AGKLMSLTAYEPSGAVALNAKLQTGAMPALKAAHQGDALFAAAYDNGYWVRLSDGGYRN
                     ASKRMLPQADKSLWSVKFAKAVSGPTAPWDKIVGQPLEIVPLEEPAAASGQIRVRVLFE
                     GRPLDGASVVATDGVNFKSEADQARATTDADGVAVVSLRSAGPQVLGVAHRVSPSQTPA
                     LADADSYGATLAFTVTDPKSN"
     gene            complement(2133..2864)
                     /gene="icuB"
                     /label=icuB
     promoter        complement(2892..2922)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     terminator      complement(3027..3054)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(3146..3232)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     complement(3414..3430)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3875..4516)
                     /codon_start=1
                     /label=TetR
                     /note="tetracycline resistance regulatory protein"
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKG"
     CDS             complement(5297..6109)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(6784..7929)
                     /codon_start=1
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     CDS             complement(8201..8572)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(8605..8714)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
ORIGIN
        1 gaccctttcc gacgctcacc gggctggttg ccctcgccgc tgggctggcg gccgtctatg
       61 gccctgcaaa cgcgccagaa acgccgtcga agccgtgtgc gagacaccgc ggccgccggc
      121 gttgtggata cctcgcggaa aacttggccc tcactgacag atgaggggcg gacgttgaca
      181 cttgaggggc cgactcaccc ggcgcggcgt tgacagatga ggggcaggct cgatttcggc
      241 cggcgacgtg gagctggcca gcctcgcaaa tcggcgaaaa cgcctgattt tacgcgagtt
      301 tcccacagat gatgtggaca agcctgggga taagtgccct gcggtattga cacttgaggg
      361 gcgcgactac tgacagatga ggggcgcgat ccttgacact tgaggggcag agtgctgaca
      421 gatgaggggc gcacctattg acatttgagg ggctgtccac aggcagaaaa tccagcattt
      481 gcaagggttt ccgcccgttt ttcggccacc gctaacctgt cttttaacct gcttttaaac
      541 caatatttat aaaccttgtt tttaaccagg gctgcgccct gtgcgcgtga ccgcgcacgc
      601 cgaagggggg tgccccccct tctcgaaccc tcccggcccg ctaacgcggg cctcccatcc
      661 ccccaggggc tgcgcccctc ggccgcgaac ggcctcaccc caaaaatggc agccaagctg
      721 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
      781 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
      841 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
      901 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
      961 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     1021 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     1081 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1141 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1201 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     1261 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     1321 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     1381 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     1441 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     1501 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     1561 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     1621 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     1681 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     1741 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     1801 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     1861 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     1921 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     1981 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     2041 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     2101 ttacgccaag cttgcatgcc taggctagcc gctcagttgg acttcgggtc ggtcacggtg
     2161 aaggcgagcg tcgcgccgta gctgtccgca tcggccagcg ccggcgtctg cgaggggctc
     2221 acgcggtgcg ccacgccgag cacctgggga ccggcactgc gcagcgagac gaccgccacg
     2281 ccgtccgcat ccgtcgtggc gcgggcctga tccgcctcgc tcttgaagtt cacgccgtcg
     2341 gtggcgacca cgctggcgcc gtcgagcgga cggccctcga acagcacccg cacccggatc
     2401 tggcccgaag ccgcggccgg ctcctccagc gggacgatct ccagcggttg gccgacgatc
     2461 ttgtcccacg gggccgtcgg gccggatacg gccttggcga acttcaccga ccagaggctc
     2521 ttgtcggcct gcggcagcat ccgcttgctg gcgttgcggt agccgccgtc ggacaagcgc
     2581 acccagtagc cgttgtcgta ggccgcggcg aacagcgcgt cgccctgatg cgcggctttg
     2641 agcgcgggca ttgcgccggt ctgcagcttg gcattcaggg ccaccgcgcc ggacggctcg
     2701 taggcggtca ggctcatcag cttgccagca ctcggcagtt ccggctcgtg gggatgcccg
     2761 tagaggatct gcacgccgtt gccggccggg tcgagccaga ggtcgtgggc ggcggccggt
     2821 gcggtgaagg cgagcagcgc ggtgaggatg acggttcgtc tcattcgggg tctctccctg
     2881 gtacctccac acaacatacg agccggaagc ataaagtgta aagcctggaa ttccctgctt
     2941 tcctgatgca aaaacgaggc tagtttaccg tatctgtggg gggatggctt gtagatatga
     3001 cgacaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttaa
     3061 tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttcg
     3121 caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac
     3181 aacagataaa acgaaaggcc cagtctttcg actgagcctt tcgttttatt tgatgcctgg
     3241 cagttcccta ctctcgcatg gggagacccc acactaccat cggcgctacg gcgtttcact
     3301 tctgagttcg gcatggggtc aggtgggacc accgcgctac tgccgccagg caaattctgt
     3361 tttatcagac cgcttctgcg ttctgattta atctgtatca ggctgaaaaa ttcactggcc
     3421 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     3481 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     3541 caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat
     3601 ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca
     3661 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     3721 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     3781 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     3841 taggttaatg tcatgataat aatggtttct tagcaccctt tctcggtcct tcaacgttcc
     3901 tgacaacgag cctccttttc gccaatccat cgacaatcac cgcgagtccc tgctcgaacg
     3961 ctgcgtccgg accggcttcg tcgaaggcgt ctatcgcggc ccgcaacagc ggcgagagcg
     4021 gagcctgttc aacggtgccg ccgcgctcgc cggcatcgct gtcgccggcc tgctcctcaa
     4081 gcacggcccc aacagtgaag tagctgattg tcatcagcgc attgacggcg tccccggccg
     4141 aaaaacccgc ctcgcagagg aagcgaagct gcgcgtcggc cgtttccatc tgcggtgcgc
     4201 ccggtcgcgt gccggcatgg atgcgcgcgc catcgcggta ggcgagcagc gcctgcctga
     4261 agctgcgggc attcccgatc agaaatgagc gccagtcgtc gtcggctctc ggcaccgaat
     4321 gcgtatgatt ctccgccagc atggcttcgg ccagtgcgtc gagcagcgcc cgcttgttcc
     4381 tgaagtgcca gtaaagcgcc ggctgctgaa cccccaaccg ttccgccagt ttgcgtgtcg
     4441 tcagaccgtc tacgccgacc tcgttcaaca ggtccagggc ggcacggatc actgtattcg
     4501 gctgcaactt tgtcatgatt gacactttat cactgataaa cataatatgt ccaccaactt
     4561 atcagtgata aagaatccgc gcgttcaatc ggaccagcgg aggctggtcc ggaggccaga
     4621 cgtgaaaccc aacatacccc tgatcgtaat tctgagcact gtcgcgctcg acgctgtcgg
     4681 catcggcctg attatgccgg tgctgccggg cctcctgcgc gatctggttc actcgaacga
     4741 cgtcaccgcc cactatggca ttctgctggc gctgtatgcg ttggtgcaat ttgcctgcgc
     4801 acctgtgctg ggcgcgctgt cggatcgttt cgggcggcgg ccaatcttgc tcgtctcgct
     4861 ggccggcgcc actgtcgact acgccatcat ggcgacagcg cctttccttt gggttctcta
     4921 tatcgggcgg atcgtggccg gcatcaccgg ggcgactggg gcggtagccg gcgcttatat
     4981 tgccgatgac ctgcaggggg gggggggcgc tgaggtctgc ctcgtgaaga aggtgttgct
     5041 gactcatacc aggcctgaat cgccccatca tccagccaga aagtgaggga gccacggttg
     5101 atgagagctt tgttgtaggt ggaccagttg gtgattttga acttttgctt tgccacggaa
     5161 cggtctgcgt tgtcgggaag atgcgtgatc tgatccttca actcagcaaa agttcgattt
     5221 attcaacaaa gccgccgtcc cgtcaagtca gcgtaatgct ctgccagtgt tacaaccaat
     5281 taaccaattc tgattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat
     5341 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac
     5401 cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa
     5461 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac
     5521 catgagtgac gactgaatcc ggtgagaatg gcaaaagctt atgcatttct ttccagactt
     5581 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat
     5641 tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac
     5701 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac
     5761 ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga
     5821 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt
     5881 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc
     5941 catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac
     6001 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg
     6061 aatttaatcg cggcctcgag caagacgttt cccgttgaat atggctcata acaccccttg
     6121 tattactgtt tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg
     6181 caatgtaaca tcagagattt tgagacacaa cgtggctttc cccccccccc ctgcaggtcc
     6241 gacacgggga tggatggcgt tcccgatcat ggtcctgctt gcttcgggtg gcatcggaat
     6301 gccggcgctg caagcaatgt tgtccaggca ggtggatgag gaacgtcagg ggcagctgca
     6361 aggctcactg gcggcgctca ccagcctgac ctcgatcgtc ggacccctcc tcttcacggc
     6421 gatctatgcg gcttctataa caacgtggaa cgggtgggca tggattgcag gcgctgccct
     6481 ctacttgctc tgcctgccgg cgctgcgtcg cgggctttgg agcggcgcag ggcaacgagc
     6541 cgatcgctga tcgtggaaac gataggccta tgccatgcgg gtcaaggcga cttccggcaa
     6601 gctatacgcg ccctagaatt gtcaatttta atcctctgtt tatcggcagt tcgtagagcg
     6661 cgccgtgcgt cccgagcgat actgagcgaa gcaagtgcgt cgagcagtgc ccgcttgttc
     6721 ctgaaatgcc agtaaagcgc tggctgctga acccccagcc ggaactgacc ccacaaggcc
     6781 ctagcgtttg caatgcacca ggtcatcatt gacccaggcg tgttccacca ggccgctgcc
     6841 tcgcaactct tcgcaggctt cgccgacctg ctcgcgccac ttcttcacgc gggtggaatc
     6901 cgatccgcac atgaggcgga aggtttccag cttgagcggg tacggctccc ggtgcgagct
     6961 gaaatagtcg aacatccgtc gggccgtcgg cgacagcttg cggtacttct cccatatgaa
     7021 tttcgtgtag tggtcgccag caaacagcac gacgatttcc tcgtcgatca ggacctggca
     7081 acgggacgtt ttcttgccac ggtccaggac gcggaagcgg tgcagcagcg acaccgattc
     7141 caggtgccca acgcggtcgg acgtgaagcc catcgccgtc gcctgtaggc gcgacaggca
     7201 ttcctcggcc ttcgtgtaat accggccatt gatcgaccag cccaggtcct ggcaaagctc
     7261 gtagaacgtg aaggtgatcg gctcgccgat aggggtgcgc ttcgcgtact ccaacacctg
     7321 ctgccacacc agttcgtcat cgtcggcccg cagctcgacg ccggtgtagg tgatcttcac
     7381 gtccttgttg acgtggaaaa tgaccttgtt ttgcagcgcc tcgcgcggga ttttcttgtt
     7441 gcgcgtggtg aacagggcag agcgggccgt gtcgtttggc atcgctcgca tcgtgtccgg
     7501 ccacggcgca atatcgaaca aggaaagctg catttccttg atctgctgct tcgtgtgttt
     7561 cagcaacgcg gcctgcttgg cctcgctgac ctgttttgcc aggtcctcgc cggcggtttt
     7621 tcgcttcttg gtcgtcatag ttcctcgcgt gtcgatggtc atcgacttcg ccaaacctgc
     7681 cgcctcctgt tcgagacgac gcgaacgctc cacggcggcc gatggcgcgg gcagggcagg
     7741 gggagccagt tgcacgctgt cgcgctcgat cttggccgta gcttgctgga ccatcgagcc
     7801 gacggactgg aaggtttcgc ggggcgcacg catgacggtg cggcttgcga tggtttcggc
     7861 atcctcggcg gaaaaccccg cgtcgatcag ttcttgcctg tatgccttcc ggtcaaacgt
     7921 ccgattcatt caccctcctt gcgggattgc cccgactcac gccggggcaa tgtgccctta
     7981 ttcctgattt gacccgcctg gtgccttggt gtccagataa tccaccttat cggcaatgaa
     8041 gtcggtcccg tagaccgtct ggccgtcctt ctcgtacttg gtattccgaa tcttgccctg
     8101 cacgaatacc agctccgcga agtcgctctt cttgatggag cgcatgggga cgtgcttggc
     8161 aatcacgcgc accccccggc cgttttagcg gctaaaaaag tcatggctct gccctcgggc
     8221 ggaccacgcc catcatgacc ttgccaagct cgtcctgctt ctcttcgatc ttcgccagca
     8281 gggcgaggat cgtggcatca ccgaaccgcg ccgtgcgcgg gtcgtcggtg agccagagtt
     8341 tcagcaggcc gcccaggcgg cccaggtcgc cattgatgcg ggccagctcg cggacgtgct
     8401 catagtccac gacgcccgtg attttgtagc cctggccgac ggccagcagg taggcctaca
     8461 ggctcatgcc ggccgccgcc gccttttcct caatcgctct tcgttcgtct ggaaggcagt
     8521 acaccttgat aggtgggctg cccttcctgg ttggcttggt ttcatcagcc atccgcttgc
     8581 cctcatctgt tacgccggcg gtagccggcc agcctcgcag agcaggattc ccgttgagca
     8641 ccgccaggtg cgaataaggg acagtgaaga aggaacaccc gctcgcgggt gggcctactt
     8701 cacctatcct gcccggctga cgccgttgga tacaccaagg aaagtctaca cgaacccttt
     8761 ggcaaaatcc tgtatatcgt gcgaaaaagg atggatatac cgaaaaaatc gctataatga
     8821 ccccgaagca gggttatgca gcggaaaaga tccgtc
//
